臺灣博碩士論文加值系統

漢氏醋桿菌(Komagataeibacter hansenii)是來自台灣當地康普茶菇菌所篩選出一種醋酸桿菌，而四季檸檬(Citruslimon (L.) osbeck)果皮是從虎尾當地市場取得。本研究以10%(w/v)四季檸檬果皮添加於10%(w/v)蔗糖水作為發酵K. hansenii所需四季檸檬皮培養基(定義為Citrus lemon sucrose, Cls)，並探討發酵時間(0~27天)對有無添加K. hansenii之Cls基質 (分別定義為ClswKh和ClswoKh)發酵上清液的 pH 值、糖度(°Brix)與抗氧化活性之影響。經過27天的K. hansenii發酵後，ClswKh上清液的pH值從開始pH3.9 緩慢下降至第27天之pH2.8，而ClswoKh上清液的pH值在儲存期間沒有變化且維持在pH4.3。在糖度方面，ClswKh與ClswoKh並未隨發酵與儲存時間而有所變化，推測K. hansenii可能利用四季檸檬果皮基質替代碳源。ClswKh上清液之抗氧化活性定義為DPPH(1,1-diphenyl 2-picrylhydrazyl)自由基清除能力(scavenging activity)(DPPH% defined as scavenging percent %)、總酚類化合物含量(total phenolic content; TPC defined as defined as galic acid equivalent, GAE)與還原能力(reducing power; RP defined as vitamin C equivalent, Vit.CE)，一般而言，ClswKh上清液之抗氧化活性皆高於ClswoKh上清液之抗氧化活性，這歸因於前培養在甘露醇基質之起始菌(starter) K. hansenii所致。ClswKh上清液之DPPH%在前9天發酵有略微上升，而後似乎持平，未有顯著變化，但其僅略高於ClswoKh上清液之DPPH%。再者，ClswKh上清液之TPC隨發酵時間增加而上升，在第15天發酵時間達到最高值0.074 mg GAE/mL，但到第18天後才逐漸下降。另外，ClswKh上清液之RP隨發酵時間增加而上升，在第12天發酵時間達到最高值0.046 Vit.CE/mL，而後逐漸下降。接著分別對ClswKh與ClswoKh上清液的DPPH%、TPC與RP值進行相互迴歸關係的探討，其中兩者的TPC-RP可得到線性關係，ClswKh線性迴歸斜率與截距值(分別接近於1.5806與0.0049 mg Vit.C/mL)略高於ClswoKh者(分別接近於1.5666與0.0030mg Vit.C/mL)，另外，對ClswKh的DPPH%-TPC迴歸關係值DPPHVmax%與1/2DPPHVmax%濃度分別接近於96.98%與0.0244 mg GAE/mL，而DPPH%-RP之DPPHVmax%與1/2DPPHVmax%濃度值分別接近於98.33%與0.0194 mg Vit.C/mL，ClswKh的迴歸關係值皆優於ClswoKh者(分別相對接近於96.53%與0.0266 mg GAE/mL和97.25%與0.0192 mg Vit.C/mL，這些迴歸關係結果顯示應該歸因於K. hansenii起始菌前培養狀態所致，而且顯示要達到相同DPPH自由基清除能力時，TPC濃度必須約1.4倍於RP濃度，因此還原力能力貢獻較多。本研究結果顯示，K. hansenii起始培養於甘露醇基質可改進提高發酵檸檬上清液的抗氧化活性能力，且貢獻於具有抗氧化活性的健康型康普檸檬之研究發展。

The 10% (w/v) peel of Citruslimon (L.) osbeck (Eureka lemon), obtained from Huwei local market, Yunlin, Taiwan, were added into 10% (w/v) sucrose as Citrus lemon sucrose (defined as Cls) for Komagataeibacter hansenii (known as one of acetic acid bacteria isolated from kombucha tea fungus) cultural medium In this study, the effect of fermentation time (0-27 days) on the pH, sugar content (oBrix), and antioxidant activity of Cls supernatant with and without K. hansenii fermentation (defined as ClswKh and ClswoKh, respectively) was investigated. The pH of ClswKh was slowly decreased with increasing fermentation time from pH 3.9 at the 0th day down to pH 2.8 at the 27th day, meanwhile those of ClswoKh were around pH 4.3 stably during the 27-day storage. Both the sugar contents of ClswKh and ClswoKh were not changed with fermentation time. The antioxidant capabilities were analyzed by DPPH (1,1-diphenyl 2-picrylhydrazyl) scavenging activity (DPPH% defined as scavenging percent %), total phenolic content (TPC defined as galic acid equivalent, GAE) and reducing power (RP defined as vitamin C equivalent, Vit.CE) assay for its activity. In general, the antioxidant activities of ClswKh were higher than those of ClswoKh which was due to K. hansenii starter precultured in the mannitol broth. The DPPH% value of ClswKh little increased with fermentation time at the first 9-day and then almost kept stably without any change. Moreover, The TPC value of ClswKh increased with fermentation time up to the highest one (0.074 mg GAE/mL) at the first 15-day and then gradually decreased from the 18th day. The RP value of ClswKh increased with fermentation time at the first 12-day and obtained the highest value 0.046 Vit.CE/mL and then gradually decreased. The regression relationships among DPPH%, TPC and RP were investigated. The linear relationships between TPC and RP for ClswKh and ClswoKh were obtained and those slope and intercept values were 1.5806 and 0.0049 mg Vit.C/mL for ClswKh, respectively, while those were 1.5666 and 0.0030mg Vit.C/mL for ClswoKh, respectively. In addition, the regression DPPHmax% and 1/2DPPHmax% values between DPPH%-TPC (96.98% and 0.0244 mg GAE/mL, respectively) and DPPH%-RP (98.33% and 0.0194 mg Vit.C/mL, respectively) for ClswKh were better than those (96.53% and 0.0266 mg GAE/mL, respectively, for DPPH%-TPC 97.25% and 0.0192 mg Vit.C/mL, respectively, for DPPH%-RP) of ClswoKh. The results of this study indicated that the concentration of TPC needs to be almost 1.4-fold of RP for the same DPPH scavenging activity (DPPH%), that is, the RP activity of ClswKh supernatant was contributed 1.4-fold higher than RP one. This study indicated the antioxidant capabilities of the Cls supernatants can be improved by the fermentation with K. hansenii to the healthy kombucha lemon with antioxidant activity.

摘要......................................................................i
Abstract..................................................................iii
誌謝......................................................................vi
目錄......................................................................vii
表目錄....................................................................ix
圖目錄....................................................................x
符號說明..................................................................xi
第一章 緒論...............................................................1
1.1 前言..................................................................1
1.2 研究目的..............................................................2
第二章 文獻回顧...........................................................3
2.1 檸檬(Citruslimon (L.) Osbeck).........................................3
2.1.1 檸檬簡介............................................................3
2.1.2 檸檬果皮及果實的生物活性成分........................................3
2.1.3 酚類化合物..........................................................6
2.2 康普茶................................................................8
2.2.1 康普茶的歷史........................................................8
2.2.2 康普茶的製造過程....................................................8
2.2.3康普茶的有益特性及化學成分...........................................10
2.2.4康普茶發酵過程中微生物的代謝.........................................12
第三章 材料與方法.........................................................13
3.1 實驗材料與儀器設備....................................................13
3.1.1 實驗儀器設備........................................................13
3.1.2 實驗材料............................................................14
3.2 發酵流程..............................................................15
3.3 分析方法..............................................................17
3.3.1 發酵液糖度測定......................................................17
3.3.2 發酵液pH值測定......................................................17
3.3.3 發酵上清液的製備....................................................17
3.3.4 還原力試驗..........................................................18
3.3.5 總酚類化合物含量試驗................................................19
3.3.6 DPPH自由基清除能力試驗..............................................20
3.4 統計分析..............................................................21
第四章 結果與討論.........................................................22
4.1 pH值及糖度變化........................................................22
4.2 DPPH自由基清除能力試驗................................................24
4.3 總酚類化合物含量試驗..................................................26
4.4 還原力能力試驗........................................................28
4.5 還原力能力、總酚類化合物含量、DPPH自由基清除能力之間的關係............30
第五章 結論...............................................................34
參考文獻...........................................................35
附錄一....................................................................45
附錄二....................................................................46
附錄三....................................................................47
附錄四....................................................................48
附錄五....................................................................49
附錄六....................................................................50
Extended Abstract.........................................................51

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